ObjectiveFor prepubertal and urgently treated malignant tumor patients, ovarian tissue cryopreservation and transplantation represent more appropriate fertility preservation methods. Current clinical practices often involve freezing ovarian tissue with high concentrations of cryoprotectants (CPAs) and thawing with water baths. These processes lead to varying degrees of toxicity and devitrification damage to ovarian tissue. Therefore, this paper proposes optimized methods for vitrification of ovarian tissues based on sodium alginate hydrogel encapsulation and magnetic induction nanowarming technology. MethodsFirstly, the study investigated the effects of sodium alginate concentration, the sequence of hydrogel encapsulation and CPAs loading on vitrification efficiency of encapsulated ovarian tissue. Additionally, the capability of sodium alginate hydrogel encapsulation to reduce the required concentration of CPAs was validated. Secondly, a platform combining water bath and magnetic induction nanowarming was established to rewarm ovarian tissue under various concentrations of magnetic nanoparticles and magnetic field strengths. The post-warming follicle survival rate, antioxidant capacity, and ovarian tissue integrity were evaluated to assess the efficacy of the method. ResultsThe study found that ovarian tissue encapsulated with 2% sodium alginate hydrogel exhibited the highest follicle survival rate after vitrification. The method of loading CPAs prior to encapsulation proved more suitable for ovarian tissue cryopreservation, effectively reducing the required concentration of CPAs by 50%. A combination of 8 g/L Fe₃O₄ nanoparticles and an alternating magnetic field of 300 Gs showed optimal warming effectiveness for ovarian tissue. Combining water bath rewarming with magnetic induction nanowarming yielded the highest follicle survival rate, enhanced antioxidant capacity, and preserved tissue morphology. ConclusionSodium alginate hydrogel encapsulation of ovarian tissue reduces the concentration of CPAs required during the freezing process. The combination of magnetic induction nanowarming with water bath provides an efficient method ovarian tissue rewarming. This study offers novel approaches to optimize ovarian tissues vitrification.

OBJECTIVE: To explore the effect of nano-hydroxyapatite/collagen composite (nHAC) on bone graft fusion after anterior cervical discectomy and fusion (ACDF) in patients with cervical spondylosis and low bone mass. METHODS: A retrospective analysis was conducted on 47 patients with low bone mass who underwent ACDF from 2017 to 2021. They were divided into the nHAC group and the allogeneic bone group according to different bone graft materials. The nHAC group included 26 cases, with 8 males and 18 females;aged 50 to 78 years old with an average of (62.81±7.79) years old;the CT value of C₂-C₇ vertebrae was (264.16±36.33) HU. The allogeneic bone group included 21 cases, with 9 males and 12 females;aged 54 to 75 years old with an average of (65.95±6.58) years old;the CT value of C₂-C₇ vertebrae was (272.39±40.44) HU. The visual analogue scale (VAS), neck disability index (NDI), and Japanese Orthopaedic Association (JOA) spinal cord function score were compared before surgery, 1 week after surgery, and at the last follow-up to evaluate the clinical efficacy. Imaging assessment included C₂-C₇ Cobb angle, surgical segment height, intervertebral fusion, and whether the cage subsidence occurred at 1 week after surgery and the last follow-up. RESULTS: The follow-up duration ranged from 26 to 39 months with an average of (33.27±3.34) months in the nHAC group and 26 to 41 months with an average of (31.86±3.57) months in the allogeneic bone group. At 1 week after surgery and the last follow-up, the VAS, NDI scores, and JOA scores in both groups were significantly improved compared with those before surgery, with statistically significant differences (P<0.05). At 1 week after surgery, the C₂-C₇ Cobb angles in the nHAC group and the allogeneic bone group were (14.26±10.32)° and (14.28±8.20)° respectively, which were significantly different from those before surgery (P<0.05). At the last follow-up, the C₂-C₇ Cobb angles in both groups were smaller than those at 1 week after surgery, with statistically significant differences (P<0.05). At 1 week after surgery, the height of the surgical segment in the nHAC group was (31.65±2.55) mm, and that in the allogeneic bone group was (33.63±3.26) mm, which were significantly different from those before surgery (P<0.05). At the last follow-up, the height of the surgical segment in both groups decreased compared with that at 1 week after surgery, with statistically significant differences (P<0.05). At the last follow-up, 39 surgical segments were fused and 6 cages subsided in the nHAC group;40 surgical segments were fused and 7 cages subsided in the allogeneic bone group;there was no statistically significant difference between the two groups (P>0.05). Compared with the CT value of vertebrae without cage subsidence, the CT value of vertebrae with cage subsidence in both groups was significantly lower, with a statistically significant difference (P<0.05). CONCLUSION The application of nHAC in ACDF for patients with low bone mass can achieve effective fusion of the surgical segment. There is no significant difference in improving clinical efficacy, intervertebral fusion, and cage subsidence compared with the allogeneic bone group. With the extension of follow-up time, the C₂-C₇ Cobb angle decreases, the height of the surgical segment is lost, and the cage subsides in both the nHAC group and the allogeneic bone group, which may be related to low bone mass. Low bone mass may be one of the risk factors for cervical spine sequence changes, surgical segment height loss, and cage subsidence after ACDF.
Humans ; Male ; Female ; Middle Aged ; Spondylosis/physiopathology* ; Spinal Fusion/methods* ; Cervical Vertebrae/surgery* ; Aged ; Diskectomy ; Durapatite ; Retrospective Studies ; Collagen/chemistry*

Gene regulation relies on the precise binding of transcription factors (TFs) at regulatory elements, but simultaneously detecting hundreds of TFs on chromatin is challenging. We developed cFOOT-seq, a cytosine deaminase-based TF footprinting assay, for high-resolution, quantitative genome-wide assessment of TF binding in both open and closed chromatin regions, even with small cell numbers. By utilizing the dsDNA deaminase SsdAtox, cFOOT-seq converts accessible cytosines to uracil while preserving genomic integrity, making it compatible with techniques like ATAC-seq for sensitive and cost-effective detection of TF occupancy at the single-molecule and single-cell level. Our approach enables the delineation of TF footprints, quantification of occupancy, and examination of chromatin influences on TF binding. Notably, cFOOT-seq, combined with FootTrack analysis, enables de novo prediction of TF binding sites and tracking of TF occupancy dynamics. We demonstrate its application in capturing cell type-specific TFs, analyzing TF dynamics during reprogramming, and revealing TF dependencies on chromatin remodelers. Overall, cFOOT-seq represents a robust approach for investigating the genome-wide dynamics of TF occupancy and elucidating the cis-regulatory architecture underlying gene regulation.
Transcription Factors/genetics* ; Humans ; Chromatin/genetics* ; Animals ; Binding Sites ; Mice ; DNA Footprinting/methods*

Objective To investigate whether microRNA-21-5p(miR-21)plays a protective role in hyperoxia-induced acute lung injury(HALI)by regulating ferroptosis through the STAT3/P53/SLC7A11 axis.Methods The interaction between STAT3 and P53 was analyzed by co-immunoprecipitation(Co-IP).Fifty 9-week-old male C57BL/6J mice were randomly divided into normoxia(Control)group,HALI group,miR-21 overexpression(miR-21)group,STAT3 inhibitor(HY-13818)group,and ferrostatin-1(Fer-1)group.After the mice from the miR-21 group received miR-21-5p AAV6 or empty vector via tracheal catheter instillation,the animals were then monitored for 3 weeks.The HY-13818 group was intraperitoneally injected with HY-13818(10 mg/kg)3 times weekly for 2 weeks.The Fer-1 group received 0.8 mg/kg ferrostatin-1 via tail vein injection once daily for 2 consecutive days during modeling.The HALI model was established by exposure to>90%oxygen.After 48 h of hyperoxia,blood samples were collected via orbital sampling for RT-PCR analysis of miR-21 expression.Lung tissues were harvested for wet/dry weight ratio and assessment of histopathological changes via HE staining for lung injury score.Activity of superoxide dismutase(SOD)and contents of malondialdehyde(MDA),Fe2?,glutathione(GSH),and reactive oxygen species(ROS)were measured using photocolorimetry,spectrophotometry and fluorometry.Western blotting was used to evaluate the protein expression of STAT3,P53,SLC7A11,and GPX4.Results The results of Co-IP showed that STAT3 could bind to P53.The HALI group exhibited obviously destroyed alveolar structure,disordered arrangement,thickened interval,with a large number of infiltrated neutrophils and collapsed alveoli,and had significantly increased pathological score of lung injury and ratio of lung Wwet/Ddry weight when compared with the Control group(P<0.05).In the miR-21 group,HY-13818 group and Fer-1 group,the severity of lung injury was significantly reduced,and the pathological score of lung injury and the ratio of Wwet/Ddry weight were decreased(P<0.05)when compared with the HALI group.Compared with the control group,the contents of MDA,Fe2+and ROS were increased(P<0.05),the activity of SOD and content of GSH were declined(P<0.05),the protein levels of STAT3 and P53 were increased(P<0.05),and those of SLC7A11 and GPX4 were decreased(P<0.05)in the HALI group.Compared with the HALI group,decreased MDA and ROS levels(P<0.05),enhanced SOD activity,Fe2+and GSH levels(P<0.05),down-regulation of STAT3 and P53(P<0.05)and up-regulation of SLC7A11 and GPX4(P<0.05)were observed in the miR-21 group and HY-13818 group.Conclusion MiR-21 alleviates HALI,which may be related to its inhibition of ferroptosis through the STAT3/P53/SLC7A11 axis.

Bone metastasis is a pathological condition in which malignant tumors originating from non-osseous tissues disseminate to bone tissue via the bloodstream,lymphatic system,or direct infiltration,inducing bone destruction and severe pain.This condition disrupts normal bone metabolism and triggers various skeletal-related events(SREs),thereby significantly impairing patients' quality of life.Current therapeutic strategies for bone metastasis include surgical intervention,radiotherapy,targeted therapy,and immunotherapy.Among these,bone-targeted therapy has shown promising potential in managing bone metastasis.Recent advancements have highlighted osteoblasts and osteoclasts,the primary regulators of bone remodeling,as critical therapeutic targets.Consequently,several bone-targeted drugs have been developed.These agents not only substantially reduce the incidence of SREs but also markedly enhance patients' quality of life and clinical outcomes.In this review,we elucidate the mechanisms of drug action targeting osteoclasts and osteoblasts,and propose potential directions for future research in bone-targeted therapy.

Objective To investigate the expression and significance of cyclin dependent kinase inhibitor 3(CDKN3)in human papillomavirus type 16(HPV16)-positive cervical cancer.Methods CDKN3 expression in pan-cancer was retrieved and downloaded from the Gene Expression Profiling Interactive Analysis(GEPIA)platform,and the expression levels of CDKN3 between normal cervical tissues(13 samples)and cervical cancer tissues(306 samples)were compared.Subsequently,GSE39001 data of HPV16-positive cervical cancer was sourced and downloaded from the Gene Expression Omnibus(GEO)database,and the expression levels of CDKN3 mRNA in HPV16-positive cervical cancer tissues(43 samples)and normal cervical tissues(12 samples)were compared.Immunohistochemical method was used to detect the expression of CDKN3 in 12 ca-ses of HPV16-positive cervical cancer,12 cases of HPV16-positive cervical precancerous lesions,10 cases of HPV16-positive chronic cervicitis and 7 cases of HPV-negative normal cervical samples collected from the Af-filiated Hospital of Guizhou Medical University.SiHa(HPV16-positive),HeLa(HPV18-positive)and HCC94(HPV-negative)cervical cancer cell lines were selected,and their CDKN3 expression were detected by West-ern blot.Results The GEPIA platform analysis showed that CDKN3 was highly expressed in pan-cancer,and the expression level of CDKN3 in cervical cancer tissue was significantly higher than that in normal cervical tissue(P<0.05).The GEO dataset reflected a significantly increased CDKN3 mRNA expression level in HPV16-positive cervical cancer compared to normal cervical tissue(P<0.001).Immunohistochemical verifi-cation showed that the positive expression rates of CDKN3 in HPV16-positive cervical cancer,HPV16-positive cervical precancerous lesion,HPV16-positive chronic cervicitis and HPV-negative normal cervical tissues were 91.7%,58.3%,0 and 0,respectively.Western blot analysis of cervical cancer cells showed that the expression level of CDKN3 in SiHa(HPV16-positive)cells was significantly higher than that in HeLa(HPV18-positive)and HCC94(HPV-negative)cells(P<0.05).Conclusion CDKN3 is a new oncogene of HPV16-positive cer-vical cancer,which may be used as a marker of cervical precancerous lesions and cervical cancer screening,and may provide a theoretical basis for subsequent mechanism research and targeted therapy.

Hydrogel is a kind of material with high water content,good biocompatibility and extracellular matrix-like property,among which polypyrrole(PPy)conductive hydrogels have both physical characteristics and excellent conductivity of hydrogels themselves.Its conductivity can be used to detect electrical signals generated in biological systems and provide electrical stimulation to regulate the activities and functions of cells and tissues.These characteristics make it widely used in the biomedical field.The recent progress of PPy conductive hydrogels in biomedical field was reviewed in this paper.In terms of classification,according to the cross-linking mechanism of PPy and hydrogel matrix,the non-covalent cross-linked PPy conductive hydrogels and covalent cross-linked PPy conductive hydrogels were divided.The applications of PPy conductive hydrogels in the biomedical field(Skin damage repair,nerve repair,myocardial repair and flexible sensing,etc.)were mainly introduced,and the development trend and challenges of PPy conductive hydrogels in the biomedical field were discussed.

Multisystem inflammatory syndrome in children(MIS-C)is a complex syndrome characterized by multi-organ involvement that has emerged in the context of the severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)outbreak.The clinical presentation of MIS-C is similar to Kawasaki disease but predominantly presents with fever and gastrointestinal symptoms,and severe cases can involve toxic shock and cardiac dysfunction.Epidemiological findings indicate that the majority of MIS-C patients test positive for SARS-CoV-2 antibodies.The pathogenesis and pathophysiology of MIS-C remain unclear,though immune dysregulation following SARS-CoV-2 infection is considered a major contributing factor.Current treatment approaches for MIS-C primarily involve intravenous immunoglobulin therapy and symptomatic supportive care.This review article provides a comprehensive overview of the definition,epidemiology,pathogenesis,clinical presentation,diagnosis,treatment,and prognosis of MIS-C.